Department of Clinical Biochemistry, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, IR, Iran.
Department of Medical Genetics, Faculty of Medicine, Iran University of Medical Sciences, Tehran, IR, Iran.
Life Sci. 2019 Sep 15;233:116682. doi: 10.1016/j.lfs.2019.116682. Epub 2019 Jul 23.
Fibrosis as the hallmark of adipose tissue dysfunction which is associated with insulin resistance and type 2 diabetes, results from deposition of excess extra cellular matrix components like collagen and increased cell death. Here we investigated the effect of antidiabetic drug, Metformin, on the factors that play role in fibrosis such as; integrin/ERK pathway, collagen VI, MMP2, MMP9, apoptosis markers including DAPK1, DAPK3, DAP, SIVA, necrosis markers including RIPK1, RIPK3, and MLKL in insulin resistant and hypertrophied adipocytes.
3T3-L1 adipocytes after differentiation to insulin resistant and hypertrophied cells, treated with Metformin, and the gene expression of aforementioned factors assayed by real time PCR. The protein expression of collagen VI and ERK assayed by western blotting.
The expression of integrins changed from 0.5 to 6-fold in hypertrophied adipocyte versus adipocyte. Apoptosis and necrosis markers increased >1.5-fold in insulin resistant and hypertrophied adipocytes. Also ECM components and ERK activation increased >2-fold and 1.5-fold, respectively in insulin resistant and hypertrophied adipocytes. Metformin caused reduction of activity of integrin/ERK pathway in Metformin treated insulin resistant and hypertrophied adipocytes compared to untreated group. Metformin also reduced collagen VI in both gene and protein expression level, MMP2 and MMP9 in gene expression, and also the expression of apoptosis and necrosis gene.
Metformin with reduction of ECM component as collagen VI, MMP2 and MMP9, integrin/ERK pathway, necrosis markers as RIPK1, RIPK3 and MLKL, and apoptosis markers including DAP, DAPK1, DAPK3 and SIVA effects on fibrosis in insulin resistant and hypertrophied adipocytes in vitro.
脂肪组织功能障碍的标志是纤维化，它与胰岛素抵抗和 2 型糖尿病有关，其原因是细胞外基质成分（如胶原）的过度沉积和细胞死亡增加。在这里，我们研究了抗糖尿病药物二甲双胍对纤维化相关因素的影响，这些因素包括整合素/ERK 通路、胶原 VI、MMP2、MMP9、凋亡标志物（包括 DAPK1、DAPK3、DAP、SIVA）、坏死标志物（包括 RIPK1、RIPK3 和 MLKL）在胰岛素抵抗和肥大脂肪细胞中的作用。
3T3-L1 脂肪细胞在分化为胰岛素抵抗和肥大细胞后，用二甲双胍处理，并通过实时 PCR 检测上述因子的基因表达。用 Western blot 法检测胶原 VI 和 ERK 的蛋白表达。
与脂肪细胞相比，肥大脂肪细胞中整合素的表达增加了 0.5 到 6 倍。凋亡和坏死标志物在胰岛素抵抗和肥大脂肪细胞中增加了 1.5 倍以上。此外，ECM 成分和 ERK 激活分别增加了 2 倍和 1.5 倍以上。与未处理组相比，二甲双胍处理的胰岛素抵抗和肥大脂肪细胞中的整合素/ERK 通路活性降低。二甲双胍还降低了胶原 VI 的基因和蛋白表达水平、MMP2 和 MMP9 的基因表达水平，以及凋亡和坏死基因的表达。
二甲双胍通过减少 ECM 成分（如胶原 VI、MMP2 和 MMP9）、整合素/ERK 通路、坏死标志物（如 RIPK1、RIPK3 和 MLKL）以及凋亡标志物（如 DAP、DAPK1、DAPK3 和 SIVA），对体外胰岛素抵抗和肥大脂肪细胞中的纤维化产生影响。
